Fig. 4

In vitro splicing results from the analysis of potential splice-altering capabilities of the c.1305 + 2 T > A and c.1305 + 2del variants. a, d Agarose gel electrophoresis of RT–PCR products of wild-type and mutated minigenes of the c.1305 + 2 T > A variant and c.1305 + 2del variant, respectively. The marker represents the DNA ladder. b, e Schematic diagram of wild-type and mutated minigene fragments. Specifically, the transcriptional mRNA sequence of wild plasmid was consistent with the expectation, including complete exon11, exon12 and exon13. The minigene plasmid expressing the c.1305 + 2 T > A variant transcribed the two aberrant transcripts: r.1305_1306ins1305 + 1_1305 + 229 and r.1305_1306ins1305 + 1_1305 + 552. The minigene plasmid expressing c.1305 + 2del transcribed the two aberrant transcripts: r.1305_1306ins1305 + 1_1305 + 228 and r.1305_1306ins1305 + 1_1305 + 551. c Sanger sequencing chromatograms of RT–PCR products of the c.1305 + 2 T > A variant. f Sanger sequencing chromatograms of RT–PCR products of the c.1305 + 2del variant